【摘要】 大肠杆菌的破碎方法对包含体的纯度有较大影响。采用酶解和超声波相结合的方式进行了大肠杆菌的破碎实验研究,考察了溶菌酶用量、酶解温度、超声处理功率和时间等因素对菌体破碎程度的影响,通过测定破碎液的A650,A280,A260nm来反映细胞的破碎程度和胞内蛋白及核酸物质的释放情况。在优化的条件下,每克湿菌体添加2mg溶菌酶,30℃酶解60min、500W超声破碎50次后,经分离洗涤可获得纯度达57%的包含体。本方法可获得纯度更高的包含体,利于重组蛋白的进一步纯化。更多还原

【Abstract】 The influence of the methods of broken E.coli on the purity of inclusion body is comparatively significant. The tests of broken E.coli were investigated by the technique of enzymolysis-ultrasound, and the influences of the addition amounts of lysozyme, the enzymolysis temperatures, the number and power of ultrasound treatment on the broken E.coli were studied. The degree of broken E.coli and the release state of proteins and nucleic acids in E.coli were revealed by A650, A280, A260 nm. Under the optimal conditions,the addition amount of lysozyme was 2 mg per gram wet cell,under 30 ℃ the enzymolysis time was 60 min, the number and power of ultrasound treatment was 50 times and 500 W. The purity of inclusion body through separated and washed can arrive to 57%. The higher purity of inclusion body can be obtained by the method. It is propitious to purify recombinant proteins.更多还原